FIG. 8.
Nuclear export: the effect of LMB on protein production. 293T cells were transfected with 1 μg of pCMV-Rev and 3 μg of pGP-RRE3, pSYNGP, or pSYNGP-RRE (with or without pCMV-Rev [1 μg]). Transfections were done in duplicate. Five hours posttransfection the medium was replaced with fresh medium in the first set and with fresh medium containing 7.5 nM LMB in the second. Twenty hours later the cells were lysed and protein production was estimated by Western blot analysis. HIV-1-positive human serum was used to detect the Gag-Pol proteins (A), and an antiactin antibody was used as an internal control (B). Lanes: 1, pGP-RRE3; 2, pGP-RRE3 plus LMB; 3, pGP-RRE3 plus pCMV-Rev; 4, pGP-RRE3 plus pCMV-Rev plus LMB; 5, pSYNGP; 6, pSYNGP plus LMB; 7, pSYNGP plus pCMV-Rev; 8, pSYNGP plus pCMV-Rev plus LMB; 9, pSYNGP-RRE; 10, pSYNGP-RRE plus LMB; 11, pSYNGP-RRE plus pCMV-Rev; 12, pSYNGP-RRE plus pCMV-Rev plus LMB.